<2023-03-28 Tue> David biweekly meeting

Reconstitution of endocytic actin network project

We've implemented the suggested changes except for the following:

  • Emily is fixing cartoon panel (work in progress, needs to add proteins)

    2023-03-28_10-23-27_2023_03_28_Selection_01.png
  • explanation of actin tail variability
    • we weren't sure if the reviewer response should go into the main text or summarized since it seems overly detailed
  • scWASP tethering explanation
    • Emily came up with a rephrasing here
    • We have data supporting that Ni-NTA/His-tag interaction is not necessary for the assay. Is this worthwhile to make a supplementary figure?
      • Emily's Western blot showing scWASP binding to beads is independent of Ni-NTA lipid

        2023-03-28_10-34-10_2023_03_28_Selection_02.png
      • my representative light microscopy data (I have lots of examples)
        • +Ni-NTA lipid

          2023-03-28_11-07-57_MAX_4.11.3M18003.jpg
          2023-03-28_11-09-50_MAX_4.11.3M18001.jpg
        • -Ni-NTA lipid

          2023-03-28_11-10-26_MAX_4.11.3M20001.jpg
          2023-03-28_11-10-45_MAX_4.11.3M20002.jpg

Mechanical role of Type I myosin in CME theory project

recent progress

  • re-installed and configured cytosim to run and analyze more simulations
  • exploring comparisons between simulations and type 1 myosin measurements 20230327_dataclub_2.png
    • model seems to predict pretty closely the optimal unbinding rate and force sensitivity for endocytic myosin

next steps

  • plot myosin step size and compare to measurements to make sure that's realistic
  • subtract each mean internalization from mean internalization without myosins to show assistive effect of myosins
    • maybe set this up by scanning number of myosins instead
  • scan a bigger parameter range; when do they become harmful to CME?
  • also scan different tensions in order to see which regime provides robustness against resistive load
    • can we predict which behavior is more assistive to mammalian-like (low load) vs. yeast-like (high load) CME?
  • generate a force vs. attachment lifetime plot for each of the simulations in order to compare to Ross's and other myo data
    • I only made this for the simulated motility assay so far, but I think it should still behave the same in the CME model
  • characterize actin organization somehow?

Date: \today

Author: Max Ferrin

Email: ferrinm@berkeley.edu

Created: 2023-03-28 Tue 13:07

Validate